Gemoll Timo, Kollbeck Sophie L, Karstens Karl F, Hò Gia G, Hartwig Sonja, Strohkamp Sarah, Schillo Katharina, Thorns Christoph, Oberländer Martina, Kalies Kathrin, Lehr Stefan, Habermann Jens K
Section for Translational Surgical Oncology and Biobanking, Department of Surgery, University of Lübeck and University Hospital Schleswig-Holstein, Campus Lübeck, D-23538 Lübeck, Germany.
Institute of Clinical Biochemistry and Pathobiochemistry, German Diabetes Center at the Heinrich-Heine-University Düsseldorf, Leibniz Center for Diabetes Research, D-40225 Düsseldorf, Germany.
Oncotarget. 2017 Jul 4;8(33):54939-54950. doi: 10.18632/oncotarget.18978. eCollection 2017 Aug 15.
While carcinogenesis in Sporadic Colorectal Cancer (SCC) has been thoroughly studied, less is known about Ulcerative Colitis associated Colorectal Cancer (UCC). This study aimed to identify and validate differentially expressed proteins between clinical samples of SCC and UCC to elucidate new insights of UCC/SCC carcinogenesis and progression.
Multiplex-fluorescence two-dimensional gel electrophoresis (2-D DIGE) and mass spectrometry identified 67 proteoforms representing 43 distinct proteins. After analysis by Ingenuity Pathway Analysis (IPA), subsequent Western blot validation proofed the differential expression of Heat shock 27 kDA protein 1 (HSPB1) and Microtubule-associated protein R/EB family, member 1 (EB1) while the latter one showed also expression differences by immunohistochemistry.
Fresh frozen tissue of UCC ( = 10) matched with SCC ( = 10) was investigated. Proteins of cancerous intestinal mucosal cells were obtained by Laser Capture Microdissection (LCM) and compared by 2-D DIGE. Significant spots were identified by mass spectrometry. After IPA, three proteins [EB1, HSPB1, and Annexin 5 (ANXA5)] were chosen for further validation by Western blotting and tissue microarray-based immunohistochemistry.
This study identified significant differences in protein expression of colorectal carcinoma cells from UCC patients compared to patients with SCC. Particularly, EB1 was validated in an independent clinical cohort.
虽然散发性结直肠癌（SCC）的致癌机制已得到充分研究，但关于溃疡性结肠炎相关结直肠癌（UCC）的了解较少。本研究旨在识别和验证SCC与UCC临床样本之间差异表达的蛋白质，以阐明UCC/SCC致癌和进展的新见解。
多重荧光二维凝胶电泳（2-D DIGE）和质谱分析鉴定出代表43种不同蛋白质的67种蛋白质异构体。经 Ingenuity 通路分析（IPA）分析后，随后的蛋白质印迹验证证实了热休克27 kDa蛋白1（HSPB1）和微管相关蛋白R/EB家族成员1（EB1）的差异表达，而后者通过免疫组织化学也显示出表达差异。
研究了与SCC（n = 10）匹配的UCC新鲜冷冻组织（n = 10）。通过激光捕获显微切割（LCM）获得癌性肠黏膜细胞的蛋白质，并通过2-D DIGE进行比较。通过质谱鉴定出显著的斑点。经IPA分析后，选择三种蛋白质[EB1、HSPB1和膜联蛋白5（ANXA5）]通过蛋白质印迹和基于组织芯片的免疫组织化学进行进一步验证。
本研究确定了UCC患者与SCC患者相比，结直肠癌细胞蛋白质表达存在显著差异。特别是，EB1在一个独立的临床队列中得到了验证。
